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Immune recognition of a human renal cancer antigen through post-translational protein splicing

Abstract

Cytotoxic T lymphocytes (CTLs) detect and destroy cells displaying class I molecules of the major histocompatibility complex (MHC) that present oligopeptides derived from aberrant self or foreign proteins. Most class I peptide ligands are created from proteins that are degraded by proteasomes and transported, by the transporter associated with antigen processing, from the cytosol into the endoplasmic reticulum, where peptides bind MHC class I molecules and are conveyed to the cell surface1. C2 CTLs, cloned from human CTLs infiltrating a renal cell carcinoma, kill cancer cells overexpressing fibroblast growth factor-5 (FGF-5)2. Here we show that C2 cells recognize human leukocyte antigen-A3 MHC class I molecules presenting a nine-residue FGF-5 peptide generated by protein splicing. This process, previously described strictly in plants3 and unicellular organisms4, entails post-translational excision of a polypeptide segment followed by ligation of the newly liberated carboxy-terminal and amino-terminal residues. The occurrence of protein splicing in vertebrates has important implications for the complexity of the vertebrate proteome and for the immune recognition of self and foreign peptides.

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Figure 1: Genetic truncation analysis of the FGF-5-encoded determinant.
Figure 2: Effect of Ala or Gly substitution on the antigenicity of the G41 construct.
Figure 3: Identification of the nonamer.
Figure 4: Determinant generation is not the result of ribosome skipping or RNA splicing.
Figure 5: Presentation of synthetic peptides by EBV-B cells.
Figure 6: HPLC analysis of synthetic and acid-stripped peptides.

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Acknowledgements

We thank J. R. Bennink and P. F. Robbins for suggestions, comments and encouragement; J. P. Riley and M. R. Parkhurst for the peptide synthesis, and S. A. Rosenberg for continuous support.

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Correspondence to Ken-ichi Hanada or James C. Yang.

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The authors declare that they have no competing financial interests.

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Hanada, Ki., Yewdell, J. & Yang, J. Immune recognition of a human renal cancer antigen through post-translational protein splicing. Nature 427, 252–256 (2004). https://doi.org/10.1038/nature02240

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